Leaf ecological stoichiometry and anatomical structural adaptation mechanisms of Quercus sect. Heterobalanus in southeastern Qinghai-Tibet Plateau.

BACKGROUND: With the dramatic uplift of the Qinghai-Tibet Plateau (QTP) and the increase in altitude in the Pliocene, the environment became dry and cold, thermophilous plants that originally inhabited ancient subtropical forest essentially disappeared. However, Quercus sect. Heterobalanus (QSH) have gradually become dominant or constructive species distributed on harsh sites in the Hengduan Mountains range in southeastern QTP, Southwest China. Ecological stoichiometry reveals the survival strategies plants adopt to adapt to changing environment by quantifying the proportions and relationships of elements in plants. Simultaneously, as the most sensitive organs of plants to their environment, the structure of leaves reflects of the long-term adaptability of plants to their surrounding environments. Therefore, ecological adaptation mechanisms related to ecological stoichiometry and leaf anatomical structure of QSH were explored. In this study, stoichiometric characteristics were determined by measuring leaf carbon (C), nitrogen (N), and phosphorus (P) contents, and morphological adaptations were determined by examining leaf anatomical traits with microscopy. RESULTS: Different QSH life forms and species had different nutrient allocation strategies. Leaves of QSH plants had higher C and P and lower N contents and higher N and lower P utilization efficiencies. According to an N: P ratio threshold, the growth of QSH species was limited by N, except that of Q. aquifolioides and Q. longispica, which was limited by both N and P. Although stoichiometric homeostasis of C, N, and P and C: N, C: P, and N: P ratios differed slightly across life forms and species, the overall degree of homeostasis was strong, with strictly homeostatic, homeostatic, and weakly homeostatic regulation. In addition, QSH leaves had compound epidermis, thick cuticle, developed palisade tissue and spongy tissue. However, leaves were relatively thin overall, possibly due to leaf leathering and lignification, which is strategy to resist stress from UV radiation, drought, and frost. Furthermore, contents of C, N, and P and stoichiometric ratios were significantly correlated with leaf anatomical traits. CONCLUSIONS: QSH adapt to the plateau environment by adjusting the content and utilization efficiencies of C, N, and P elements. Strong stoichiometric homeostasis of QSH was likely a strategy to mitigate nutrient limitation. The unique leaf structure of the compound epidermis, thick cuticle, well-developed palisade tissue and spongy tissue is another adaptive mechanism for QSH to survive in the plateau environment. The anatomical adaptations and nutrient utilization strategies of QSH may have coevolved during long-term succession over millions of years.

Efflux and assimilation of xylem-transported CO2 in stems and leaves of tree species with different wood anatomy.

Determining the fate of CO2 respired in woody tissues is necessary to understand plant respiratory physiology and to evaluate CO2 recycling mechanisms. An aqueous 13 C-enriched CO2 solution was infused into the stem of 3-4 m tall trees to estimate efflux and assimilation of xylem-transported CO2 via cavity ring-down laser spectroscopy and isotope ratio mass spectrometry, respectively. Different tree locations (lower stem, upper stem and leafy shoots) and tissues (xylem, bark and leaves) were monitored in species with tracheid, diffuse- and ring-porous wood anatomy (cedar, maple and oak, respectively). Radial xylem CO2 diffusivity and xylem [CO2 ] were lower in cedar relative to maple and oak trees, thereby limiting label diffusion. Part of the labeled 13 CO2 was assimilated in cedar (8.7%) and oak (20.6%) trees, mostly in xylem and bark tissues of the stem, while limited solution uptake in maple trees hindered the detection of label assimilation. Little label reached foliar tissues, suggesting substantial label loss along the stem-branch transition following reductions in the radial diffusive pathway. Differences in respiration rates and radial xylem CO2 diffusivity (lower in conifer relative to angiosperm species) might reconcile discrepancies in efflux and assimilation of xylem-transported CO2 so far observed between taxonomic clades.

Atmospheric pCO2 impacts leaf structural and physiological traits in Quercus petraea seedlings.

MAIN CONCLUSION: Atmospheric p CO 2 impacts Quercus petraea biomass production and cell wall composition of the leaves in favor of cellulose at the expense of lignin, and enhances foliar non-structural carbohydrate levels and sucrose contents in a pCO 2 concentration-dependent manner. Sessile oak (Quercus petraea Liebl.) was grown for ca. half a year from seeds at ambient control (525 ppm), 750, 900, and 1000 ppm atmospheric pCO2 under controlled conditions. Increasing pCO2 enhanced biomass production, modified the cell wall composition of the leaves in favor of cellulose at the expense of lignin, and enhanced the foliar non-structural carbohydrate level, in particular the sucrose content; as well as total N content of leaves by increased levels of all major N fractions, i.e., soluble proteins, total amino acids, and structural N. The enhanced total amino acid level was largely due to 2-ketoglutarate and oxalo acetate-derived compounds. Increasing pCO2 alleviated oxidative stress in the leaves as indicated by reduced H2O2 contents. High in vitro glutathione reductase activity at reduced H2O2 contents suggests enhanced ROS scavenging, but increased lipid peroxidation may also have contributed, as indicated by a negative correlation between malone dialdehyde and H2O2 contents. Almost all these effects were at least partially reversed, when pCO2 exceeded 750 or 900 ppm. Apparently, the interaction of atmospheric pCO2 with leaf structural and physiological traits of Q. petraea seedlings is characterized by a dynamic response depending on the pCO2 level.

Influence of dry season on Quercus suber L. leaf traits in the Iberian Peninsula.

PREMISE: Water deficit and drought conditions are increasing in intensity, frequency, and duration in the Iberian Peninsula. We observed natural variation in leaf traits across the range of Quercus suber L. (cork oak), an ecologically important species within this region. Stomatal traits (e.g., pore length, maximum aperture) and carbon isotope composition (δ13 C) provide an opportunity to examine the integrative effects of drought and dry-season intensity on leaf development, maximum stomatal conductance, and adaptation to precipitation regimes. METHODS: Gross leaf traits (e.g., area, thickness), stomatal traits (e.g., pore length, size, aperture), and carbon isotope discrimination were measured in Q. suber leaves, and maximum stomatal conductance to water vapor (gwmax ) was calculated. Trees were sampled from nine natural populations across a climate gradient in the Iberian Peninsula, including trees from two genetic lineages. Linear mixed models compared total water deficit to leaf traits, accounting for tree and site as random effects. RESULTS: Quercus suber gross leaf morphology remained consistent across the climate gradient, but increasing water deficit was correlated with smaller stomata at the leaf level and low δ13 C at the tree level. No traits were significantly different between the two genetic lineages. CONCLUSIONS: While there were no significant differences in gross leaf morphology across the climate gradient or between the genetic lineages, stomatal traits and δ13 C responded to climate, suggesting that Q. suber can inhabit a range of environments in the Iberian Peninsula via micro-adjustments of the trait that controls water loss into the atmosphere.

Immobilisation of yeasts on oak chips or cellulose powder for use in bottle-fermented sparkling wine.

Sparkling wine production comprises two successive fermentations performed by Sacharomyces cerevisiae strains. This research aimed to: develop yeast immobilisation processes on two wine-compatible supports; study the effects of yeast type (IOC 18-2007 and 55A) and the immobilisation support type (oak chips and cellulose powder) on the fermentation kinetics, the deposition rate of lees and the volatile composition of the finished sparkling wine; compare the fermentation parameters of the wines inoculated with immobilised or non-immobilised cells. Proper immobilisation of yeast on oak chips and cellulose powder was demonstrated by electron microscopy. Total sugar consumption occurred in under 60 days in all bottles, regardless of the strain used and the way they were inoculated in wine. Deposition of lees was 3-fold faster in the bottles containing immobilised cells than in those with free cells; no addition of adjuvants was necessary. The analysis of the volatile compounds of the finished sparkling wines showed significant differences in the formation of esters, acids, alcohols, aldehydes and lactones according to the yeast and the immobilisation support used. Oak chips were the more appropriate support for yeast immobilisation. No significant differences in the sensorial analysis of the sparkling wines produced by the different strategies were found.

Natural selection and neutral evolutionary processes contribute to genetic divergence in leaf traits across a precipitation gradient in the tropical oak Quercus oleoides.

The impacts of drought are expanding worldwide as a consequence of climate change. However, there is still little knowledge of how species respond to long-term selection in seasonally dry ecosystems. In this study, we used QST -FST comparisons to investigate (i) the role of natural selection on population genetic differentiation for a set of functional traits related to drought resistance in the seasonally dry tropical oak Quercus oleoides and (ii) the influence of water availability at the site of population origin and in experimental treatments on patterns of trait divergence. We conducted a thorough phenotypic characterization of 1912 seedlings from ten populations growing in field and greenhouse common gardens under replicated watering treatments. We also genotyped 218 individuals from the same set of populations using eleven nuclear microsatellites. QST distributions for leaf lamina area, specific leaf area, leaf thickness and stomatal pore index were higher than FST distribution. Results were consistent across growth environments. Genetic differentiation among populations for these functional traits was associated with the index of moisture at the origin of the populations. Together, our results suggest that drought is an important selective agent for Q. oleoides and that differences in length and severity of the dry season have driven the evolution of genetic differences in functional traits.

Linking fine root morphology, hydraulic functioning and shade tolerance of trees.

Background and Aims: Understanding root traits and their trade-off with other plant processes is important for understanding plant functioning in natural ecosystems as well as agricultural systems. The aim of the present study was to determine the relationship between root morphology and the hydraulic characteristics of several orders of fine roots (<2 mm) for species differing in shade tolerance (low, moderate and high). Methods: The morphological, anatomical and hydraulic traits across five distal root orders were measured in species with different levels of shade tolerance and life history strategies. The species studied were Acer negundo, Acer rubrum, Acer saccharum, Betula alleghaniensis, Betula lenta, Quercus alba, Quercus rubra, Pinus strobus and Pinus virginiana. Key Results: Compared with shade-tolerant species, shade-intolerant species produced thinner absorptive roots with smaller xylem lumen diameters and underwent secondary development less frequently, suggesting that they had shorter life spans. Shade-tolerant species had greater root specific hydraulic conductance among these roots due to having larger diameter xylems, although these roots had a lower calculated critical tension for conduit collapse. In addition, shade-intolerant species exhibited greater variation in hydraulic conductance across different root growth rings in woody transport roots of the same root order as compared with shade-tolerant species. Conclusions: Plant growth strategies were extended to include root hydraulic properties. It was found that shade intolerance in trees was associated with conservative root hydraulics but greater plasticity in number of xylem conduits and hydraulic conductance. Root traits of shade-intolerant species were consistent with the ability to proliferate roots quickly for rapid water uptake needed to support rapid shoot growth, while minimizing risk in uncertain environments.

Is stemflow a vector for the transport of small metazoans from tree surfaces down to soil?

BACKGROUND: Stemflow is an essential hydrologic process shaping the soil of forests by providing a concentrated input of rainwater and solutions. However, the transport of metazoans by stemflow has yet to be investigated. This 8-week study documented the organisms (< 2 mm) present in the stemflow of different tree species. Because the texture of the tree bark is a crucial determination of stemflow, trees with smooth bark (Carpinus betulus and Fagus sylvatica) and rough bark (Quercus robur) were examined. RESULTS: Up to 1170 individuals per liter of stemflow were collected. For rotifers and nematodes, a highly positive correlation between abundance and stemflow yield was determined. Both taxa were predominant (rotifers: up to 70%, nematodes: up to 13.5%) in the stemflow of smooth-barked trees whereas in that of the oak trees collembolans were the most abundant organisms (77.3%). The mean number of organisms collected per liter of stemflow from the two species of smooth-barked trees was very similar. A higher number of nematode species was found in the stemflow of these trees than in the stemflow of rough-barked oak and all were typical colonizers of soil- and bark-associated habitats. CONCLUSION: This pilot study showed for the first time that stemflow is a transport vector for numerous small metazoans. By connecting tree habitats (e.g., bark, moss, lichens or water-filled tree holes) with soil, stemflow may influence the composition of soil fauna by mediating intensive organismal dispersal.

Noninvasive Measurement of Vulnerability to Drought-Induced Embolism by X-Ray Microtomography.

Hydraulic failure induced by xylem embolism is one of the primary mechanisms of plant dieback during drought. However, many of the methods used to evaluate the vulnerability of different species to drought-induced embolism are indirect and invasive, increasing the possibility that measurement artifacts may occur. Here, we utilize x-ray computed microtomography (microCT) to directly visualize embolism formation in the xylem of living, intact plants with contrasting wood anatomy (Quercus robur, Populus tremula × Populus alba, and Pinus pinaster). These observations were compared with widely used centrifuge techniques that require destructive sampling. MicroCT imaging provided detailed spatial information regarding the dimensions and functional status of xylem conduits during dehydration. Vulnerability curves based on microCT observations of intact plants closely matched curves based on the centrifuge technique for species with short vessels (P. tremula × P. alba) or tracheids (P. pinaster). For ring porous Q. robur, the centrifuge technique significantly overestimated vulnerability to embolism, indicating that caution should be used when applying this technique to species with long vessels. These findings confirm that microCT can be used to assess the vulnerability to embolism on intact plants by direct visualization.

Stem hydraulic capacitance decreases with drought stress: implications for modelling tree hydraulics in the Mediterranean oak Quercus ilex.

Hydraulic modelling is a primary tool to predict plant performance in future drier scenarios. However, as most tree models are validated under non-stress conditions, they may fail when water becomes limiting. To simulate tree hydraulic functioning under moist and dry conditions, the current version of a water flow and storage mechanistic model was further developed by implementing equations that describe variation in xylem hydraulic resistance (RX ) and stem hydraulic capacitance (CS ) with predawn water potential (ΨPD ). The model was applied in a Mediterranean forest experiencing intense summer drought, where six Quercus ilex trees were instrumented to monitor stem diameter variations and sap flow, concurrently with measurements of predawn and midday leaf water potential. Best model performance was observed when CS was allowed to decrease with decreasing ΨPD . Hydraulic capacitance decreased from 62 to 25 kg m-3 MPa-1 across the growing season. In parallel, tree transpiration decreased to a greater extent than the capacitive water release and the contribution of stored water to transpiration increased from 2.0 to 5.1%. Our results demonstrate the importance of stored water and seasonality in CS for tree hydraulic functioning, and they suggest that CS should be considered to predict the drought response of trees with models.

Evolutionary dynamics of the leaf phenological cycle in an oak metapopulation along an elevation gradient.

It has been predicted that environmental changes will radically alter the selective pressures on phenological traits. Long-lived species, such as trees, will be particularly affected, as they may need to undergo major adaptive change over only one or a few generations. The traits describing the annual life cycle of trees are generally highly evolvable, but nothing is known about the strength of their genetic correlations. Tight correlations can impose strong evolutionary constraints, potentially hampering the adaptation of multivariate phenological phenotypes. In this study, we investigated the evolutionary, genetic and environmental components of the timing of leaf unfolding and senescence within an oak metapopulation along an elevation gradient. Population divergence, estimated from in situ and common-garden data, was compared to expectations under neutral evolution, based on microsatellite markers. This approach made it possible (1) to evaluate the influence of genetic correlation on multivariate local adaptation to elevation and (2) to identify traits probably exposed to past selective pressures due to the colder climate at high elevation. The genetic correlation was positive but very weak, indicating that genetic constraints did not shape the local adaptation pattern for leaf phenology. Both spring and fall (leaf unfolding and senescence, respectively) phenology timings were involved in local adaptation, but leaf unfolding was probably the trait most exposed to climate change-induced selection. Our data indicated that genetic variation makes a much smaller contribution to adaptation than the considerable plastic variation displayed by a tree during its lifetime. The evolutionary potential of leaf phenology is, therefore, probably not the most critical aspect for short-term population survival in a changing climate.

Do changes in spring phenology affect earlywood vessels? Perspective from the xylogenesis monitoring of two sympatric ring-porous oaks.

This study addresses relationships between leaf phenology, xylogenesis, and functional xylem anatomy in two ring-porous oak species, the temperate Quercus robur and the sub-Mediterranean Q. pyrenaica. Earlywood vessel (EV) formation and leaf phenology were monitored in 2012 and 2013. Ten individuals per species were sampled at each of three sites located in NW Iberian Peninsula. EV areas measured on microcore sections were used to calculate the hydraulic tree diameter (Dh ), in order to model relationships to phenology. Thermal requirements were evaluated using growing degree days (GDD). A species-specific timing of growth resumption was found. The onset of EV formation and budburst were associated to a particular GDD in each species. The onset and duration of EV enlargement affected Dh (and EV size) in Q. robur, but hardly in Q. pyrenaica. The relationship between the timings of EV formation and xylem structure appears to be stronger for the temperate oak, whose larger vessels may result from thermal-induced earlier resumption. In contrast, the sub-Mediterranean oak would maintain a more conservative hydraulic architecture under warming conditions.

Pure species in a continuum of genetic and morphological variation: sympatric oaks at the edge of their range.

BACKGROUND AND AIMS: Studies on oaks (Quercus spp.) have often been hampered by taxonomic confusion, a situation further compounded by the occurrence of extensive interspecific hybridization. In the present study, a combination of genetic and morphological analyses was used to examine sympatric populations of Q. petraea and Q. robur at the north-western edge of their ranges in Northern Ireland, since it had previously been suggested that hybridization could facilitate the apparent rapid, long-distance dispersal of oaks following the glaciations. METHODS: Samples were collected from 24 sites across Northern Ireland that had been previously designated as ancient or semi-natural woodland. Genotypes were obtained from a total of 950 trees using 12 nuclear microsatellite loci, and admixture coefficients were calculated based on a Bayesian clustering approach. Individuals were also classified as Q. petraea,Q. robur or hybrids based on two objective morphometric characters shown previously to delineate pure individuals effectively. Genetically 'pure' individuals of both species, as defined by the Bayesian clustering, were also genotyped for five chloroplast microsatellites. KEY RESULTS: Genetic and morphological analyses both indicated the presence of pure individuals of both species, as well as a continuum of intermediates. There was a good agreement between the molecular and morphological classification, with a generally clear separation between pure individuals. CONCLUSIONS: Despite millennia of hybridization and introgression, genetically and morphologically pure individuals of both Q. petraea and Q. robur can be found at the edge of their range, where both species occur sympatrically. The high proportion of individuals exhibiting introgression compared with previous studies may reflect the historical role of hybridization in facilitating dispersal following the glaciations. This is further supported by the significantly higher chloroplast diversity in Q. robur compared with Q. petraea.

A field-to-desktop toolchain for X-ray CT densitometry enables tree ring analysis.

BACKGROUND AND AIMS: Disentangling tree growth requires more than ring width data only. Densitometry is considered a valuable proxy, yet laborious wood sample preparation and lack of dedicated software limit the widespread use of density profiling for tree ring analysis. An X-ray computed tomography-based toolchain of tree increment cores is presented, which results in profile data sets suitable for visual exploration as well as density-based pattern matching. METHODS: Two temperate (Quercus petraea, Fagus sylvatica) and one tropical species (Terminalia superba) were used for density profiling using an X-ray computed tomography facility with custom-made sample holders and dedicated processing software. KEY RESULTS: Density-based pattern matching is developed and able to detect anomalies in ring series that can be corrected via interactive software. CONCLUSIONS: A digital workflow allows generation of structure-corrected profiles of large sets of cores in a short time span that provide sufficient intra-annual density information for tree ring analysis. Furthermore, visual exploration of such data sets is of high value. The dated profiles can be used for high-resolution chronologies and also offer opportunities for fast screening of lesser studied tropical tree species.

Bidirectional anatomical effects in a mistletoe-host relationship: Psittacanthus schiedeanus mistletoe and its hosts Liquidambar styraciflua and Quercus germana.

PREMISE OF THE STUDY: During the interactions between a parasitic plant and its host, the parasite affects its host morphologically, anatomically, and physiologically, yet there has been little focus on the effect of hosts on the parasite. Here, the functional interactions between the hemiparasitic mistletoe Psittacanthus schiedeanus and its hosts Liquidambar styraciflua and Quercus germana were interpreted based on the anatomical features of the vascular tissues. METHODS: Using standard techniques for light and transmission electron microscopy, we studied the effects of P. schiedeanus on the phloem anatomy of Liquidambar styraciflua and Quercus germana and vice versa. KEY RESULTS: The phloem of P. schiedeanus has larger sieve elements, companion cells, and sieve plate areas when it is parasitizing L. styraciflua than Q. germana; however, the parasite produces systemic effects on the phloem of its hosts, reducing the size of phloem in L. styraciflua but increasing it in Q. germana. Those seem to be the bidirectional effects. No direct connections between the secondary phloem of the parasite and that of its hosts were observed. Parenchymatic cells of L. styraciflua in contact with connective parenchyma cells of the parasite develop half-plasmodesmata, while those of Q. germana do not. CONCLUSIONS: The bidirectional effects between the parasite and its hosts comprise modifications in secondary phloem that are potentially affected by the phenology of its hosts, a combination of hormonal agents such as auxins, and the symplasmic or apoplasmic pathway for solutes import.

Losses of leaf area owing to herbivory and early senescence in three tree species along a winter temperature gradient.

In temperate climates, evergreen leaves have to survive throughout low temperature winter periods. Freezing and chilling injuries can lead to accelerated senescence of part of the leaf surface, which contributes to a reduction of the lifespan of the photosynthetic machinery and of leaf lifetime carbon gain. Low temperatures are also associated with changes in foliar chemistry and morphology that affect consumption by herbivores. Therefore, the severity of foliar area losses caused by accelerated senescence and herbivory can change along winter temperature gradients. The aim of this study is to analyse such responses in the leaves of three evergreen species (Quercus ilex, Q. suber and Pinus pinaster) along a climatic gradient. The leaves of all three species presented increased leaf mass per area (LMA) and higher concentrations of structural carbohydrates in cooler areas. Only the two oak species showed visible symptoms of damage caused by herbivory, this being less intense at the coldest sites. The leaves of all three species presented chlorotic and necrotic spots that increased in size with leaf age. The foliar surface affected by chlorosis and necrosis was larger at the sites with the coldest winters. Therefore, the effects of the winter cold on the lifespan of the photosynthetic machinery were contradictory: losses of leaf area due to accelerated senescence increased, but there was a decrease in losses caused by herbivory. The final consequences for carbon assimilation strongly depend on the exact timing of the appearance of the damage resulting from low temperature and grazing by herbivores.

A comparison of two methods for measuring vessel length in woody plants.

Vessel lengths are important to plant hydraulic studies, but are not often reported because of the time required to obtain measurements. This paper compares the fast dynamic method (air injection method) with the slower but traditional static method (rubber injection method). Our hypothesis was that the dynamic method should yield a larger mean vessel length than the static method. Vessel length was measured by both methods in current year stems of Acer, Populus, Vitis and Quercus representing short- to long-vessel species. The hypothesis was verified. The reason for the consistently larger values of vessel length is because the dynamic method measures air flow rates in cut open vessels. The Hagen-Poiseuille law predicts that the air flow rate should depend on the product of number of cut open vessels times the fourth power of vessel diameter. An argument is advanced that the dynamic method is more appropriate because it measures the length of the vessels that contribute most to hydraulic flow. If all vessels had the same vessel length distribution regardless of diameter, then both methods should yield the same average length. This supports the hypothesis that large-diameter vessels might be longer than short-diameter vessels in most species.

A new positive relationship between pCO2 and stomatal frequency in Quercus guyavifolia (Fagaceae): a potential proxy for palaeo-CO2 levels.

BACKGROUND AND AIMS: The inverse relationship between atmospheric CO2 partial pressure (pCO2) and stomatal frequency in many species of plants has been widely used to estimate palaeoatmospheric CO2 (palaeo-CO2) levels; however, the results obtained have been quite variable. This study attempts to find a potential new proxy for palaeo-CO2 levels by analysing stomatal frequency in Quercus guyavifolia (Q. guajavifolia, Fagaceae), an extant dominant species of sclerophyllous forests in the Himalayas with abundant fossil relatives. METHODS: Stomatal frequency was analysed for extant samples of Q. guyavifolia collected from17 field sites at altitudes ranging between 2493 and 4497 m. Herbarium specimens collected between 1926 and 2011 were also examined. Correlations of pCO2-stomatal frequency were determined using samples from both sources, and these were then applied to Q. preguyavaefolia fossils in order to estimate palaeo-CO2 concentrations for two late-Pliocene floras in south-western China. KEY RESULTS: In contrast to the negative correlations detected for most other species that have been studied, a positive correlation between pCO2 and stomatal frequency was determined in Q. guyavifolia sampled from both extant field collections and historical herbarium specimens. Palaeo-CO2 concentrations were estimated to be approx. 180-240 ppm in the late Pliocene, which is consistent with most other previous estimates. CONCLUSIONS: A new positive relationship between pCO2 and stomatal frequency in Q. guyavifolia is presented, which can be applied to the fossils closely related to this species that are widely distributed in the late-Cenozoic strata in order to estimate palaeo-CO2 concentrations. The results show that it is valid to use a positive relationship to estimate palaeo-CO2 concentrations, and the study adds to the variety of stomatal density/index relationships that available for estimating pCO2. The physiological mechanisms underlying this positive response are unclear, however, and require further research.

[Identification of Epmedii Folium and its counterfeit leaf of Quercus variabilis].

Epmedii Folium is a commonly used traditional Chinese drug, and is beneficial for the "liver" and "kidney" s function in Chinese medicine. Recently, the origin of this drug is more complex. Most of the identification studies are emphasized on the species certified by the pharmacopoeia and other related species from the same genus of Epimedium, but few was emphasized on the counterfeit. In this paper, one counterfeit of Epmedii Folium, identified as the dried leaf of Quercus variabilis (Fam. Fagaceae), has been reported based on field investigation, comparing specimen of Epmedii Folium and Q. variabilis,using the macroscopic, microscopic and TmC methods. It is resulted that they could be identified clearly not only by the macroscopic features, such as the vein character and the tooth apex, but also by the microscopic features, such as the vascular bundles of the midrib, the non-glandular hair, the anticlinal wall of the epidermis cell and the calcium oxalate crystal. Furthermore their TLC chromatograms showed also difference. This study will give reference for the identification of Epmedii Folium and the related supervision and inspection work.

Stem girdling affects the quantity of CO2 transported in xylem as well as CO2 efflux from soil.

There is recent clear evidence that an important fraction of root-respired CO2 is transported upward in the transpiration stream in tree stems rather than fluxing to the soil. In this study, we aimed to quantify the contribution of root-respired CO2 to both soil CO2 efflux and xylem CO2 transport by manipulating the autotrophic component of belowground respiration. We compared soil CO2 efflux and the flux of root-respired CO2 transported in the transpiration stream in girdled and nongirdled 9-yr-old oak trees (Quercus robur) to assess the impact of a change in the autotrophic component of belowground respiration on both CO2 fluxes. Stem girdling decreased xylem CO2 concentration, indicating that belowground respiration contributes to the aboveground transport of internal CO2 . Girdling also decreased soil CO2 efflux. These results confirmed that root respiration contributes to xylem CO2 transport and that failure to account for this flux results in inaccurate estimates of belowground respiration when efflux-based methods are used. This research adds to the growing body of evidence that efflux-based measurements of belowground respiration underestimate autotrophic contributions.